Silke Smolders1, Sofie Kessels2, Sophie Marie-Thérèse Smolders3, Florent Poulhes4, Olivier Zelphati5, Cedric Sapet6, Bert Brône7. 1. UHasselt, BIOMED, Diepenbeek, Belgium; Laboratory of Neuronal Differentiation, VIB Center for the Biology of Disease, Leuven and Center for Human Genetics, KU Leuven, Leuven, Belgium. Electronic address: silke.smolders@uhasselt.be. 2. UHasselt, BIOMED, Diepenbeek, Belgium. Electronic address: sofie.kessels@uhasselt.be. 3. UHasselt, BIOMED, Diepenbeek, Belgium; Inserm, Umr-s 1130, Cnrs, Umr 8246, Neuroscience Paris Seine, Institute of Biology Paris Seine, Paris, France; Sorbonne Universités, Upmc Université Paris 06, Um Cr18, Neuroscience Paris Seine, Paris, France. Electronic address: sophie.smolders@uhasselt.be. 4. Oz Biosciences,parc Scientifique De Luminy - Zone Entreprise. 163 Avenue De Luminy, Case 922. 13288 Marseille Cedex 9, France. Electronic address: fpoulhes@ozbiosciences.com. 5. Oz Biosciences,parc Scientifique De Luminy - Zone Entreprise. 163 Avenue De Luminy, Case 922. 13288 Marseille Cedex 9, France. Electronic address: ozelphati@ozbiosciences.com. 6. Oz Biosciences,parc Scientifique De Luminy - Zone Entreprise. 163 Avenue De Luminy, Case 922. 13288 Marseille Cedex 9, France. Electronic address: csapet@ozbiosciences.com. 7. UHasselt, BIOMED, Diepenbeek, Belgium. Electronic address: Bert.brone@uhasselt.be.
Abstract
BACKGROUND: Microglia, the resident phagocytic cells of the brain, have recently been the subject of intense investigation given their role in pathology and normal brain physiology. In general, phagocytic cells are hard to transfect with plasmid DNA. The BV2 cell line is a murine cell line of microglial origin which is often used to study this cell type in vitro. Unfortunately, this microglial cell line is, like other phagocytic cells, resistant to transfection. NEW METHOD: Magnetofection is a well-established transfection method that combines DNA with magnetic particles which, under the influence of a magnetic field, ensures a high concentration of particles in proximity of cultured cells. Only recently, Glial-Mag was specifically developed for efficient transfection of microglia and microglial cell lines. RESULTS: Magnetofection with Glial-Mag yielded a transfection efficiency of 34.95% in BV2 cells, 24h after transfection with an eGFP-expressing plasmid. Efficient gene delivery caused a modest and short-lived cell activation (as measured by IL6 secretion) that ceased by 24h after transfection. COMPARISON WITH EXISTING METHODS: Here we show that Glial-Mag magnetofection of BV2 cells yielded a significantly higher transfection efficiency (34.95%) compared to other chemical transfection methods including calcium-phoshate precipication (0.34%), X-tremeGENE (3.30%) and Lipofectamine 2000 (12.51%). CONCLUSION: Transfection of BV2 cells using Glial-Mag magnetofection is superior compared to other chemical transfection methods and could be considered as the method of choice to chemically transfect microglial cell lines.
BACKGROUND: Microglia, the resident phagocytic cells of the brain, have recently been the subject of intense investigation given their role in pathology and normal brain physiology. In general, phagocytic cells are hard to transfect with plasmid DNA. The BV2 cell line is a murine cell line of microglial origin which is often used to study this cell type in vitro. Unfortunately, this microglial cell line is, like other phagocytic cells, resistant to transfection. NEW METHOD: Magnetofection is a well-established transfection method that combines DNA with magnetic particles which, under the influence of a magnetic field, ensures a high concentration of particles in proximity of cultured cells. Only recently, Glial-Mag was specifically developed for efficient transfection of microglia and microglial cell lines. RESULTS: Magnetofection with Glial-Mag yielded a transfection efficiency of 34.95% in BV2 cells, 24h after transfection with an eGFP-expressing plasmid. Efficient gene delivery caused a modest and short-lived cell activation (as measured by IL6 secretion) that ceased by 24h after transfection. COMPARISON WITH EXISTING METHODS: Here we show that Glial-Mag magnetofection of BV2 cells yielded a significantly higher transfection efficiency (34.95%) compared to other chemical transfection methods including calcium-phoshate precipication (0.34%), X-tremeGENE (3.30%) and Lipofectamine 2000 (12.51%). CONCLUSION: Transfection of BV2 cells using Glial-Mag magnetofection is superior compared to other chemical transfection methods and could be considered as the method of choice to chemically transfect microglial cell lines.